| 质粒名称: | LeGO-lnc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80624 |
| 存储实验室: | Jan-Henning Klusmann |
|
载体骨架基本信息 |
|
| 载体名称: | LeGO-Cer |
| 载体出品公司: | Kristoffer Ricken, Boris Fehse |
| 空载体大小: | 8300 bp |
| 完整质粒大小: | 8300 bp |
| 载体修饰: | 删除mU6启动子，插入带有lncRNA MCS（PacI，SgsI）的克隆寡核苷酸，插入BGHpolyA，插入PGK启动子 |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | 杀稻瘟菌素,麦克鲁林 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | empty control |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | SFFV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SgsI（销毁后未知） |
| 3’克隆位点: | PacI（销毁后未知） |
| 5’测序引物 | GAGCTCACAACCCCTCACTC |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | LeGO-lnc was a gift from Jan-Henning Klusmann (Addgene plasmid # 80624 ; http://n2t.net/addgene:80624 ; RRID:Addgene_80624) |
| 参考文献部分: | LincRNAs MONC and MIR100HG act as oncogenes in acute megakaryoblastic leukemia. Emmrich S, Streltsov A, Schmidt F, Thangapandi VR, Reinhardt D, Klusmann JH. Mol Cancer. 2014 Jul 15;13:171. doi: 10.1186/1476-4598-13-171. 10.1186/1476-4598-13-171 PubMed 25027842 |
买家导航
LeGOlnc载体基本信息
LeGOlnc载体图谱质粒图谱和多克隆位点信息
LeGOlnc载体简介
LeGOlnc载体序列
LOCUS LeGO-lnc 8301 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral lncRNA Overexpression.
ACCESSION .
VERSION .
KEYWORDS LeGO-lnc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8301)
AUTHORS Emmrich S, Streltsov A, Schmidt F, Thangapandi VR, Reinhardt D,
Klusmann JH
TITLE LincRNAs MONC and MIR100HG act as oncogenes in acute
megakaryoblastic leukemia.
JOURNAL Mol Cancer. 2014 Jul 15;13:171. doi: 10.1186/1476-4598-13-171.
PUBMED 25027842
REFERENCE 2 (bases 1 to 8301)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8301
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2104..2148
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2446..2563
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2622..2638
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2680..2713
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal 3205..3429
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 3436..3934
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(3470..3493)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3867..3886
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 3952..3961
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3958..4674
/codon_start=1
/product="enhanced monomeric variant of CFP (Rizzo et al.,
2004)"
/label=mCerulean
/note=" mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4003..4024)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4264..4283)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4611..4632
/label=EGFP-C
/note="EGFP, forward primer"
CDS 4675..5070
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
AIVKDSDGQPTAVGIRELLPSGYVWEG"
protein_bind 5101..5134
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 5190..5778
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5243..5263)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5661..5672)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5781..5797)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 6307..6487
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6549..7137)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6629..6648)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7308..8168)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7931..7950
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8169..8273)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
2581 cagcagagat ccagtttggt tagtaccggg cccgctctag tcgaggtcga cggtatcgat
2641 aagctcgctt cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat
2701 tatacgaagt tatattaagg gttccaagct taagcggccg ctgaaagacc ccacctgtag
2761 gtttggcaag ctagctgcag taacgccatt ttgcaaggca tggaaaaata ccaaaccaag
2821 aatagagaag ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg gccaaacagg
2881 atatctgcgg tgagcagttt cggccccggc ccggggccaa gaacagatgg tcaccgcagt
2941 ttcggccccg gcccgaggcc aagaacagat ggtccccaga tatggcccaa ccctcagcag
3001 tttcttaaga cccatcagat gtttccaggc tcccccaagg acctgaaatg accctgcgcc
3061 ttatttgaat taaccaatca gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga
3121 gctctataaa agagctcaca acccctcact cggcgcgcca gtcctccgac ctaggtctag
3181 actcgagtta taattaatta acgactgtgc cttctagttg ccagccatct gttgtttgcc
3241 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
3301 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
3361 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
3421 gctctatgga tgcatggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
3481 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
3541 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
3601 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
3661 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
3721 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
3781 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
3841 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
3901 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccggatcc cgccaccatg
3961 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
4021 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
4081 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
4141 gtgaccaccc tgacctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
4201 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
4261 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
4321 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
4381 ctggagtaca acgccatcag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc
4441 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
4501 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
4561 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
4621 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggccaag
4681 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
4741 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
4801 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
4861 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
4921 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
4981 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
5041 ctgccctctg gttatgtgtg ggagggctaa tgtacaagta agaattcgtc gagggaccta
5101 ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact
5161 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
5221 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
5281 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
5341 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
5401 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
5461 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
5521 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
5581 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
5641 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
5701 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
5761 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga
5821 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
5881 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
5941 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
6001 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
6061 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
6121 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
6181 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
6241 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac
6301 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
6361 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
6421 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
6481 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
6541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
6601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
6661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
6721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
6781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
6841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
6901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
6961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
7021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
7081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
7141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
7201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
7261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
7321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
7381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
7441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
7501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
7561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
7621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
7681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
7741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
7801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
7861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
7921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
7981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
8041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
8101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
8161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
8221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
8281 ccccgaaaag tgccacctga c
//
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